... Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...to studying membranes, organelles, and other subcellular compartments is via proteomics--the study of ...promiscuous labeling enzyme and a small-molecule substrate to map the spatiotemporal distribution and interaction...sequencing, revealing that they bind to a much larger subset of protein motifs than previously thought. Sidhu...

...your experiment. To make an FMO sample, you take a subset of your experimental samples, combine it into one...be from the same host species, have the same Ig subclass, and be in the same fluorophore (e.g., if you ...staining for anti-mouse Ly6C in APC with the IgG2c subclass, you will use the APC IgG2c isotype control). ...a FMO sample: you make a sample from a combined subset of your experimental samples, stain it with every...

...empirically determined. In addition, some metal ion substrates, such as those containing Co2+, have a lower ...affinity for polyhistidine than more commonly used substrates such as Ni2+ and are therefore more selective... systems, consider using these more selective substrates along with stringent wash conditions.   Find ...tightly the protein will bind to the metal ion substrate. If your protein of interest is expressed at a...

...Applications (RFA), what next? First, check the submission deadline and try not to panic if it's due in ...in mind that universities require application submission in-house approximately 2 weeks before the RFA..., award information, eligibility information, submission information, selection criteria, and award administration...written grant application builds upon the last submission. Thus, continuing to apply for funding improves...

... molecule, such as biotin. Next, a conjugated substrate is added to the wells. In the case where the detection...enzyme horseradish peroxidase (HRP). Finally, the substrate of HRP is added to the wells. This reaction produces...detected by an enzyme-conjugated antibody. Next, its substrate is added and the reaction can be visualized via...that results from the reaction of HRP with its substrate, 3,3′,5,5′-Tetramethylbenzidine (TMB). The addition...

...biotinylated target protein can be recovered for the subsequent steps. Phage display mRNA from the enriched pool...transcribed into cDNA fragments that are efficiently subcloned into the phagemid pDX_init plasmid, using the ...sybody pools from the phage display screens are subcloned (using FX cloning) into the expression vector ...for identification. All unique sybodies can be subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for...

...Cas6. Regardless of the subtype, the mechanism of Cascade binding and subsequent DNA degradation appears... These type Is can then be further broken into subtypes of A–G, which come from different species and ...and type IV with an unknown effector. Type I has subtypes A through G, with different compositions of the...

...larger than the substance P peptide, but still quite small. However, unlike the substance P tag, they found...experiment involved a peptide from the neuropeptide substance P. This protein had a highly specific antibody...meaning the tag could be quite small. However, substance P has an amide group added to its C-terminus post-translationally...

...BiFC studies used the method to interrogate the subcellular localization of interactions between basic leucine...fusion proteins it requires can lead to aberrant subcellular localization of the studied PIPs. The straightforward...endogenous counterparts using western blotting. The subcellular localization of BiFC constructs should also be...because their expression level, stability, and subcellular localization may differ from those of your BiFC...

...synthesizing fluorophore fusions on their own. Subsequent advances in fluorescent proteins have replicated...from a sample. Chemical fluorophores can be substantially brighter and more photostable than fluorescent...reacts specifically and irreversibly with a small substrate ligand motif attached to a fluorophore (Figure...incorporation of four-membered azetidine rings could substantially enhance the brightness and photostability of...

...CamR gene. Screening for surviving colonies and subsequent rounds of evolution and engineering gave rise...TadA*) that satisfactorily accepts DNA as its substrate (see figure 2).          The artificially evolved...which is treated as ‘G’ by cellular polymerases. Subsequently, an original genomic A-T base pair is converted...stable transformation, regenerated plants may be subjected to genotyping. If successful base editing causes...

...Every few months, we highlight a subset of the new plasmids, antibodies, and viral preps in the repository...compartments. They showed APEX2 can use an orthogonal substrate, alkyne-phenol, to label proteins with an alkyne...development in and of itself. After adding one enzyme’s substrate, then the other, a dual-enrichment and mass spectrometry...Andrews, N.P., et al. (2019). A toolbox of IgG subclass-switched recombinant monoclonal antibodies for...

...produce substances toxic to themselves? What are toxin-antitoxin systems? These toxic substances are part...inferes with DNA replication by binding the GyrA subunit of DNA gyrase and trapping it in the cleaved complex... in vitro without antibiotic selection. In a subcutaneous cancer model, the researchers showed that after...

.... In the specific example of episodic memory, a subset of hippocampal neurons projects axons directly ...tracing has the added benefit of only labelling the subset of infected cells that also express Cre. Another...the ability to spread trans-synaptically on to subsequent neurons. AAV vectors are replication incompetent...incompetent and cannot readily be passed on to subsequent cells via replication like RABV or HSV. There are,...transsynaptic transfer of a protein, tetanus toxin, subsequent to its retrograde axonal transport. J Cell Biol...

...can be used to introduce specific nucleotide substitutions (or deletions) in a tailored manner. The approach...perform alanine scanning mutagenesis or targeted substitution of key residues), and in SNP analysis (to introduce...site through mutagenesis vastly facilitates the subsequent process of screening for succesfully mutated ... difficult to sequence), it may be easier to subsequently clone a smaller region (containing the specific...

...group as well as evolution of the coelenterzine substrate to the more stable yet brighter furimazine by ...i.p. or i.v. administration of the furimazine substrate for NanoLuc luciferase. Both GpNLuc and OgNLuc...multiple cellular events such as the dynamics of subcellular structures and gene expression. You must see ...sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem Biol 7, 1848-57. PubMed PMID: 22894855...

...Anyone can submit data from their CLOVER lab or Addgene AAV experiments to the Data Hub. Submit your data...Interested? You can learn more here - and remember, submissions are due Nov. 21!   ...

...example, cell sorting. The separated cells are subsequently sequenced by next generation sequencing to identify...approach is to use smaller libraries targeting subsets of genes. With such an approach, you lose the unbiased...stimulation would then be separated by sorting, and subsequently sequenced to identify which signaling molecules...Inc for this purpose.  The gRNA oligos are subsequently cloned into your vector of choice using the ...

...as they cast their ballots for their favorite submission. So without any further ado, we present to you...receives an honorable mention for their second submission.    Our Cultured Warrior (made by Nate using ...from Lab Support at the John Innes Centre, and submitted by the Lab Support Team at the Centre.     Lab...

...of GFP, its brighter mutant S65T-GFP, and the subsequent biochemical characterization, has enabled scientists...mitochondrial targeting sequence of cytochrome c oxidase subunit VIII, so that they can be imported to the mitochondrial...Chlamydomonas noctigama (CnChR1), Chloromonas subdivisa (CsChR), Chloromonas oogama (CoChR), and Scherffelia...electron microscopy (EM) tag providing superior subcellular localization of tagged proteins compared to traditional...proteins of interest for EM imaging, or targeted to subcellular regions or protein complexes for proteomic mapping...